酿酒酵母菌及其灭活菌对绵羊瘤胃外植体β-防御素-1(SBD-1)表达的影响

旨在探索益生性酿酒酵母菌及其灭活菌对绵羊瘤胃外植体内β-防御素-1(sheep beta-defensin-1,SBD-1)表达的影响。本研究建立了绵羊瘤胃外植体培养方法,将不同浓度(10~4、10~5、10~6、10~7、10~8、10~9 CFU·mL^-1)的酿酒酵母菌及其灭活菌与外植体共培养24 h后,通过qPCR和ELISA方法检测瘤胃外植体内SBD-1 mRNA和蛋白的表达变化,以分别确定活菌及其灭活菌诱导SBD-1表达最高的菌液浓度;然后用该浓度的活菌及其灭活菌对瘤胃外植体进行不同时间(2、4、8、12、16、20、24 h)的刺激,同样用qPCR和ELISA方法检测瘤胃外植体内SBD-1 mRNA及蛋白的表达变化,从而筛选出活菌及其灭活菌诱导SBD-1表达的最佳时间。结果表明,益生性酿酒酵母菌及其灭活菌均能显著促进绵羊瘤胃外植体SBD-1的表达(P<0.05)。当酿酒酵母菌浓度为10~7 CFU·mL^-1、灭活菌浓度为10~8 CFU·mL^-1分别诱导绵羊瘤胃外植体16 h时,SBD-1表达量分别达到最大,且二者与对照相比均呈极显著差异(P<0.01)。在最佳诱导条件下对比二者的诱导效果,酿酒酵母菌活菌高于其灭活菌。因此,益生性酿酒酵母菌及其灭活菌均能促进绵羊瘤胃外植体内SBD-1的表达,且活菌浓度为10~7 CFU·mL^-1、灭活菌浓度为10~8 CFU·mL^-1分别诱导16 h时,绵羊瘤胃外植体内的SBD-1的表达量分别达到最大,并且益生性酿酒酵母菌的诱导效果高于其灭活菌。     

TGF-β家族成员调控家禽卵泡发育的研究进展及展望

与哺乳动物不同,家禽的卵泡发育无明显发情周期,其卵泡发育遵循严格的等级发育体系。禽类卵泡的等级发育是一个复杂的调节过程,受卵泡细胞内外的内分泌激素、自分泌/旁分泌因子的共同调节。促卵泡刺激素(Follicle stimulating hormone,FSH)和促黄体激素(Luteinizing hormone,LH)在调控卵泡发育、卵母细胞成熟和排卵中起着重要的作用。此外,局部产生的生长因子如转化生长因子β超家族(Transforming growth factor superfamily,TGF-β)成员,包括抗缪勒氏激素(Anti-Müllerian hormone,AMH)、骨形态蛋白家族(Bone morphogenetic proteins,BMPs)、抑制素(Inhibin)和激活素(Activin)等,与促性腺激素一起协同调控卵泡的生长发育、卵母细胞的成熟等过程。文章综述了TGF-β家族成员,包括AMH、BMPs、抑制素、激活素在家禽卵泡发育中的作用及其未来的研究方向。     

过瘤胃氯化胆碱在奶牛上的应用研究进展

高产奶牛在产后泌乳初期体内常出现能量供应负平衡状态,继而引发如脂肪肝、酮病等一系列营养代谢性疾病。在产后奶牛日粮中外源适量添加过瘤胃氯化胆碱(rumen-protected choline, PRC)能改善血液生化指标,提高奶牛生产性能。作者综合了过瘤胃氯化胆碱的有关文章和试验,简要综述了其在奶牛上应用的研究进展。     

《园艺学报》征稿简则

AIM:To investigate the prognostic value of biomarkers on predicting recurrence of hepatocellular carcinoma (HCC) after orthotopic liver transplantation (OLT). METHODS:Fifty patients with HCC who underwent OLT between April 2002 and November 2005 with a minimum clinical follow up of 12 months were included in this retrospective study. We examined immunohistochemical expression of E-cadherin with β-catenin, Ki-67 proliferative index and analyzed the correlation between these biomarkers with recurrence and survival, along with the main clinical-pathological variables.RESULTS:Only at univariate analysis, TNM, portal vein tumor thrombi were valuable on predicting recurrence and survival time (P<0.05), and preoperative serum AFP correlate to recurrence only at multivariate analysis (OR=2.552, P<0.05). Lower membrane expression of E-cadherin, nuclear β-catenin localization and high Ki-67 index showed notable significance on predicting recurrence and survival time at univariate analysis, as well at multivariate analysis, all P<0.01. The membrane expression of β-catenin did not correlate to the prognosis (P>0.05).CONCLUSION:These three biomarkers might have potential as a tumor prognostic marker for predicting recurrence of HCC after OLT and perhaps are better than the clinical-pathological variables.     

β-丙氨酸的生理功能及其在动物生产中的应用

β-丙氨酸是一种不参与蛋白质合成的氨基酸,为肌肽、鹅肌肽等肌源活性肽合成的前体物,作为增强肌肉耐力的运动营养补充剂已经广泛应用于临床营养。研究表明,β-丙氨酸可提高动物生产性能,调控肌肉生长和肌源活性肽含量,改善肉品质量。本文旨在对β-丙氨酸的来源和代谢、生理功能及其在动物生产领域的应用进行综述,为β-丙氨酸在动物营养调控和生产实践中的应用提供理论依据。     

β-葡聚糖对肉仔鸡生长性能、免疫功能和肠道微环境的影响

本试验旨在研究饲粮中添加β-葡聚糖对肉仔鸡生长性能、免疫功能和肠道微环境的影响。选取1日龄爱拔益加(AA)肉仔鸡672只,随机分为4个组,每组14个重复,每个重复12只鸡。对照组饲喂基础饲粮,试验组分别饲喂在基础饲粮上添加100、150和200 g/tβ-葡聚糖的试验饲粮。试验期为42 d。结果表明:1)150 g/t剂量组1~21日龄肉仔鸡体增重显著高于对照组(P0.05)。2)150 g/t剂量组肉仔鸡血清免疫球蛋白含量高于对照组(P0.05)。3)21日龄时,150 g/t剂量组肉仔鸡盲肠内乳酸菌数量显著高于对照组(P0.05),空肠和回肠内沙门氏菌数量显著低于对照组(P0.05);42日龄时,150和200 g/t剂量组肉仔鸡盲肠内沙门氏菌数量显著低于对照组和100 g/t剂量组(P0.05)。4)饲粮添加β-葡聚糖对42日龄肉仔鸡空肠绒毛高度与隐窝深度比值影响显著(P0.05)。由此可见,饲粮中添加适量的β-葡聚糖可提高肉仔鸡出栏重,改善肉仔鸡生长性能,增加盲肠内乳酸菌数量,减少空肠、回肠和盲肠内沙门氏菌数量。     

鸡β-actin基因实时荧光定量PCR方法的建立

根据GenBank上鸡β-actin基因的序列，在保守区域设计并合成一对引物，采用SYBR Green I染料建立了荧光定量PCR法（real-time PCR）。以常规PCR产物为标准品，建立了标准曲线，并进行了熔解曲线分析。结果表明，标准曲线Ct值检测范围为12~31,扩增效率为95.1%；熔解曲线分析结果显示产物特异的单个峰，其Tm为88±0℃，检测周期从RNA提取到荧光定量PCR结束只需4 h。本试验建立的鸡β-actin基因实时荧光定量PCR法扩增效率高、检测范围广、检测周期短，为β-actin基因作为内参基因进行鸡功能基因与病原基因表达的定量分析奠定了基础。     

Synthetic tetramer of a Phytophthora sojae-inducible fragment from soybean GmaSKTI36 promoter improves its pathogen induction activities

Using pathogen-induced promoters to control expression of the functional genes in transgenic plants may greatly increase the chances of boosting disease resistance. However, the number of the inducible promoters is limited. Here, we found that soybean GmaSKTI36 gene is strongly induced upon Phytophthora sojae infection. Functional analysis showed that its promoter could mediate rapid and strong induction of GUS expression upon pathogen infection in both Nicotiana benthamiana leaves and soybean hairy roots. Then, a 122 bp fragment that was critical to the activity was successfully identified by a progressive 5′ deletion analysis. Importantly, we found that a synthetic promoter by tetramerizing this fragment could confer strong P. sojae induction activities. Overall, the results suggested that the GmaSKTI36 promoter, the 122 bp fragment, and the synthetic promoter are potentially useful pathogen-inducible promoters.     

Short-term dietary supplementation with the microalga Parietochloris incisa enhances stress resistance in guppies Poecilia reticulata

Two trials were conducted to determine the effects of dietary enrichments with the microalga Parietochloris incisa , rich in arachidonic acid (ARA), on stress resistance in guppies Poecilia reticulata . The microalga was added to commercial diets as a neutral lipid (NL) extract and its fractions or as broken cells. Experimental diets were applied for a period of 14 days. In trial 1, commercial diets were supplemented with NL (containing 25 mg ARA and 0.11 mg β-carotene g⁻¹ feed), its triacylglycerol (TAG) fraction (containing 25 mg ARA g⁻¹ feed and no β-carotene) and the β-carotene fraction (containing 0.11 mg carotenoid g⁻¹ feed and minute amounts of ARA). Neutral lipid-fed fish demonstrated the highest resistance ( P <0.05) to osmotic stress (32-ppt NaCl), followed by fish fed with diets supplemented with TAG and β-carotene alone, which were more resistant than control ( P <0.05). In trial 2, fish fed diets supplemented with higher levels of broken alga (26.1 mg ARA g⁻¹ feed) were more resistant ( P <0.05) to stress as compared with fish fed lower ARA (16.3 mg g g⁻¹) or an unsupplemented control diet. We suggest a dietary supplementation with broken P. incisa cells to enhance stress resistance in guppies before a stressful event.     

TGF-β/Smad信号通路响应光周期变化参与调控斑马鱼卵巢发育

为研究不同光周期条件下转化生长因子-β超家族/Samd (TGF-β/Smad)信号通路在斑马鱼(Danio rerio)卵巢中的作用,利用荧光定量PCR技术检测连续黑暗(0 L∶24 D, DD)、自然光照(14 L∶10 D, LD)和连续光照(24 L∶0 D, LL)条件下斑马鱼卵巢中TGF-β/Smad信号通路中相关基因的相对表达量,并利用免疫组化技术检测不同光周期条件下p-Smad2在斑马鱼卵巢中的定位情况。结果显示,在不同光周期条件下,TGF-β/Smad信号通路中信号分子在斑马鱼卵巢中的表达模式不尽相同,其中配体(tgfb3)、受体(tgfbr2a,tgfbr2b,tgfbr1b)和下游蛋白激酶(smad2和smad3a)的表达趋势一致,即在不同光周期处理3 d后,上述6个基因的相对表达量DD组最高,LL组最低,但处理7 d后则呈相反趋势。在斑马鱼卵黄发生前期至充分生长未成熟期的卵母细胞中均检测到p-Smad2蛋白信号,但光周期处理后未影响p-Samd2在斑马鱼卵巢中的定位。结果表明,光周期变化可能通过改变tgfb3/受体/蛋白激酶基因的表达模式影响斑马鱼的卵巢发育。